Partial correction of endogenous ΔF508 CFTR in human cystic fibrosis airway epithelia by spliceosome-mediated RNA trans-splicing

X. Liu, Q. Jiang, S. G. Mansfield, M. Puttaraju, Y. Zhang, W. Zhou, J. A. Cohn, Mariano Garcia-Blanco, L. G. Mitchell, J. F. Engelhardt

Research output: Contribution to journalArticle

127 Citations (Scopus)

Abstract

Spliceosome-mediated RNA trans-splicing (SMaRT) was investigated as a means for functionally correcting endogenous ΔF508 cystic fibrosis transmembrane conductance regulator (CFTR) transcripts using in vitro human cystic fibrosis (CF) polarized airway epithelia and in vivo human CF bronchial xenografts. Recombinant adenovirus (Ad.CFTR-PTM) encoding a pre-therapeutic molecule (PTM) targeted to CFTR intron 9 corrected transepithelial cyclic AMP (cAMP)-sensitive short-circuit current (Isc) in ΔF508 homozygous epithelia to a level 16% of that observed in normal human bronchial epithelia. Molecular analyses using RT-PCR and western blotting confirmed SMaRT-mediated partial correction of endogenous ΔF508 messenger RNA (mRNA) transcripts and protein. In an in vivo model of ΔF508 CF airway epithelia, human CF bronchial xenografts infected with Ad.CFTR-PTM also demonstrated partial correction of CFTR-mediated Cl- permeability at a level 22% of that seen in non-CF xenografts. These results provide functional evidence for SMaRT-mediated repair of mutant endogenous CFTR mRNA in intact polarized CF airway epithelial models.

Original languageEnglish (US)
Pages (from-to)47-52
Number of pages6
JournalNature Biotechnology
Volume20
Issue number1
DOIs
StatePublished - 2002
Externally publishedYes

Fingerprint

Trans-Splicing
Spliceosomes
Cystic Fibrosis Transmembrane Conductance Regulator
RNA
Cystic Fibrosis
Epithelium
Molecules
Heterografts
Short circuit currents
Repair
Proteins
Messenger RNA
Adenoviridae
Cyclic AMP
Introns
Permeability
Fibrosis
Therapeutics
Western Blotting
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Microbiology

Cite this

Liu, X., Jiang, Q., Mansfield, S. G., Puttaraju, M., Zhang, Y., Zhou, W., ... Engelhardt, J. F. (2002). Partial correction of endogenous ΔF508 CFTR in human cystic fibrosis airway epithelia by spliceosome-mediated RNA trans-splicing. Nature Biotechnology, 20(1), 47-52. https://doi.org/10.1038/nbt0102-47

Partial correction of endogenous ΔF508 CFTR in human cystic fibrosis airway epithelia by spliceosome-mediated RNA trans-splicing. / Liu, X.; Jiang, Q.; Mansfield, S. G.; Puttaraju, M.; Zhang, Y.; Zhou, W.; Cohn, J. A.; Garcia-Blanco, Mariano; Mitchell, L. G.; Engelhardt, J. F.

In: Nature Biotechnology, Vol. 20, No. 1, 2002, p. 47-52.

Research output: Contribution to journalArticle

Liu, X, Jiang, Q, Mansfield, SG, Puttaraju, M, Zhang, Y, Zhou, W, Cohn, JA, Garcia-Blanco, M, Mitchell, LG & Engelhardt, JF 2002, 'Partial correction of endogenous ΔF508 CFTR in human cystic fibrosis airway epithelia by spliceosome-mediated RNA trans-splicing', Nature Biotechnology, vol. 20, no. 1, pp. 47-52. https://doi.org/10.1038/nbt0102-47
Liu, X. ; Jiang, Q. ; Mansfield, S. G. ; Puttaraju, M. ; Zhang, Y. ; Zhou, W. ; Cohn, J. A. ; Garcia-Blanco, Mariano ; Mitchell, L. G. ; Engelhardt, J. F. / Partial correction of endogenous ΔF508 CFTR in human cystic fibrosis airway epithelia by spliceosome-mediated RNA trans-splicing. In: Nature Biotechnology. 2002 ; Vol. 20, No. 1. pp. 47-52.
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